肾透明细胞癌中VHL基因失活的检测及意义

目的 探讨肾透明细胞癌中Von Hippel-Lindau(VHL)基因突变和超甲基化情况及其在肾癌发生发展中的作用及临床意义.方法 采用聚合酶链反应-单链构象多态性分析(PCRSSCP)银染法,甲基化特异性PCR(MSP)及测序等方法对33例肾透明细胞癌和相应远离肿瘤正常肾组织VHL基因突变和超甲基化情况进行检测.结果 19/33(57.6%)例肾透明细胞癌中出现VHL基因突变;6/33(18.2%)例肾透明细胞癌中出现VHL基因超甲基化;VHL基因突变和超甲基化与肾癌临床分期和淋巴结转移相关.结论 肾癌中存在VHL基因突变和超甲基化,且与临床分期、淋巴结转移相关;VHL基因失活的检测可作为肾透明细胞癌的诊断指标,VHL基因可望成为肾透明细胞癌基因治疗的重要目的 基因.     

von Hipple-Lindau基因突变与散发性肾透明细胞癌患者预后的关系

目的 探讨von Hippel-Lindau(VHL)抑癌基因突变与散发性肾透明细胞癌患者预后的关系. 方法应用聚合酶链反应(PCR)、PCR产物直接测序分析74例散发性肾透明细胞癌组织标本和相应远离肿瘤的正常组织标本中VHL基因突变情况.74例患者,病理分期为T1 51例(68.9%),T2 9例(12.2%),T3 14例(18.9%);病理分级为G1 15例(20.3%),G2 50例(67.6%),G39例(12.2%).随访其预后并进行统计学分析. 结果 VHL基因突变者40例(54.1%),不同病理分期和分级的肿瘤中VHL基因突变率差异无统计学意义(P值分别为0.915和0.237).随访34~107个月.平均71个月,因肿瘤死亡7例,出现远处肿瘤转移11例,5年无瘤生存率为78%.VHL基因突变组肿瘤死亡或转移等阳性事件发生率(15.0%,6/40)明显低于非突变组(35.3%,12/34,P=0.043).Logistic回归分析表明.患者预后与肿瘤病理分期、分级呈正相关,而与VHL基因是否突变呈负相关,三者的P值分别为0.016、0.024和0.033.对于T3和G3肿瘤患者,VHL基因突变者预后更好.P值分别为0.010和0.048. 结论 散发性肾透明细胞癌患者中VHL基因突变广泛,肾癌的病理分期和分级仍然是评估患者预后的有效指标.VHL基因突变失活可能提示肾透明细胞癌患者预后更好,尤其对于高分期和高分级肿瘤患者.     

散发性肾透明细胞癌组织中希佩尔-林道基因突变与缺氧诱导因子1α、2α的测定

目的探讨散发性肾透明细胞癌(CCRCC)组织中希佩尔-林道(VHL)基因突变、缺氧诱导因子1α(HIF-1α)和HIF-2α的表达及其关系,对肿瘤分期、分级的影响.方法应用聚合酶链反应(PCR)、PCR产物直接测序和免疫组化等方法,分析77例散发性CCRCC患者癌组织中VHL基因突变、HIF-1α和HIF-2α的表达.其中T1期 55例(71%),T2期7例(9%),T3期14例(18%),T4期1例(1%);病理分级,G115例(19%),G256例(73%),G36例(8%).结果在正常肾组织中无VHL基因突变.散发性CCRCC中VHL基因突变率为52%(40/77),HIF-2α阳性率81% (62/77),高于HIF-1α阳性率66%(51/77)(χ2 =23.310, P<0.01);VHL基因突变者中HIF-1α和HIF-2α的阳性率(98%和93%)均高于无突变者的阳性率(32% 和68%,χ2值分别为36.386,7.617,P均<0.01);HIF-1α和HIF-2α的表达均与VHL基因突变相关(偏回归系数分别为4.481,2.027,P均<0.01);未发现VHL基因突变、HIF-1α和HIF-2α表达与肿瘤病理分级、分期有关(P均>0.05).结论在散发性CCRCC患者中VHL基因突变较广泛,在突变组织中HIF-1α和2α高表达,但VHL基因突变、HIF-1α和2α的表达与患者病理分级、分期不相关.     

肾细胞癌患者抑癌基因VHL双等位基因

目的探讨肾细胞癌患者中抑癌基因VHL的双等位基因失活发生情况.方法在41例肾细胞癌患者中提取肿瘤和正常组织DNA.采用单链聚合酶链反应(PCR)和测序法检测肿瘤组织中VHL基因的突变情况.采用PCR限制性片段长度多态性法检测VHL基因内部的2个单核苷酸多态性(SNP)位点,在2个位点的杂合子中通过对比肿瘤组织和正常肾组织分析VHL基因的杂合性丢失(LOH)情况.结果在肾细胞癌中51%(21/41)发生VHL基因突变,42%(8/19)发生VHL基因LOH,基因突变和LOH发生具有显著一致性(r=0.78),在37%(7/19)的肾细胞癌中发生VHL双等位基因失活.结论肾细胞癌中存在由VHL基因突变和LOH导致的VHL双等位基因失活现象,VHL双等位基因失活发生率为37%.     

肾细胞癌与VHL 基因

肾细胞癌是泌尿生殖系统最重要的肿瘤之一.其中约有4%为遗传性肾癌.肾癌中最常见的病理类型为透明细胞性肾细胞癌.几乎所有的遗传性肾癌和绝大多数的散发性透明细胞性肾细胞癌源于VHL 基因的失活.本综述主要关注于肾细胞癌和VHL 基因之间的联系.涉及肾细胞癌的遗传学分型、可能的发病机制、HIF 依赖途径的 VHL 功能、HIF 非依赖途径的 VHL 功能以及 VHL 基因失活的肾细胞患者预后.同时介绍针对VHL 其下游基因的分子靶向治疗药物及相关疗效.     

肾细胞癌与VHL 基因

肾细胞癌是泌尿生殖系统最重要的肿瘤之一.其中约有4%为遗传性肾癌.肾癌中最常见的病理类型为透明细胞性肾细胞癌.几乎所有的遗传性肾癌和绝大多数的散发性透明细胞性肾细胞癌源于VHL 基因的失活.本综述主要关注于肾细胞癌和VHL 基因之间的联系.涉及肾细胞癌的遗传学分型、可能的发病机制、HIF 依赖途径的 VHL 功能、HIF 非依赖途径的 VHL 功能以及 VHL 基因失活的肾细胞患者预后.同时介绍针对VHL 其下游基因的分子靶向治疗药物及相关疗效.     

肾细胞癌与VHL 基因

肾细胞癌是泌尿生殖系统最重要的肿瘤之一.其中约有4%为遗传性肾癌.肾癌中最常见的病理类型为透明细胞性肾细胞癌.几乎所有的遗传性肾癌和绝大多数的散发性透明细胞性肾细胞癌源于VHL 基因的失活.本综述主要关注于肾细胞癌和VHL 基因之间的联系.涉及肾细胞癌的遗传学分型、可能的发病机制、HIF 依赖途径的 VHL 功能、HIF 非依赖途径的 VHL 功能以及 VHL 基因失活的肾细胞患者预后.同时介绍针对VHL 其下游基因的分子靶向治疗药物及相关疗效.     

肾细胞癌与VHL 基因

肾细胞癌是泌尿生殖系统最重要的肿瘤之一.其中约有4%为遗传性肾癌.肾癌中最常见的病理类型为透明细胞性肾细胞癌.几乎所有的遗传性肾癌和绝大多数的散发性透明细胞性肾细胞癌源于VHL 基因的失活.本综述主要关注于肾细胞癌和VHL 基因之间的联系.涉及肾细胞癌的遗传学分型、可能的发病机制、HIF 依赖途径的 VHL 功能、HIF 非依赖途径的 VHL 功能以及 VHL 基因失活的肾细胞患者预后.同时介绍针对VHL 其下游基因的分子靶向治疗药物及相关疗效.     

肾细胞癌与VHL 基因

肾细胞癌是泌尿生殖系统最重要的肿瘤之一.其中约有4%为遗传性肾癌.肾癌中最常见的病理类型为透明细胞性肾细胞癌.几乎所有的遗传性肾癌和绝大多数的散发性透明细胞性肾细胞癌源于VHL 基因的失活.本综述主要关注于肾细胞癌和VHL 基因之间的联系.涉及肾细胞癌的遗传学分型、可能的发病机制、HIF 依赖途径的 VHL 功能、HIF 非依赖途径的 VHL 功能以及 VHL 基因失活的肾细胞患者预后.同时介绍针对VHL 其下游基因的分子靶向治疗药物及相关疗效.     

肾细胞癌与VHL 基因

肾细胞癌是泌尿生殖系统最重要的肿瘤之一.其中约有4%为遗传性肾癌.肾癌中最常见的病理类型为透明细胞性肾细胞癌.几乎所有的遗传性肾癌和绝大多数的散发性透明细胞性肾细胞癌源于VHL 基因的失活.本综述主要关注于肾细胞癌和VHL 基因之间的联系.涉及肾细胞癌的遗传学分型、可能的发病机制、HIF 依赖途径的 VHL 功能、HIF 非依赖途径的 VHL 功能以及 VHL 基因失活的肾细胞患者预后.同时介绍针对VHL 其下游基因的分子靶向治疗药物及相关疗效.     

Nuclear expression of hypoxia-inducible factor-1alpha in clear cell renal cell carcinoma is involved in tumor progression

OBJECTIVES: The most frequent genomic abnormality in clear cell renal cell carcinoma (cc-RCC) is inactivation of Von Hippel-Lindau gene (VHL). pVHL19 is a ligase promoting proteosomal degradation of hypoxia-inducible factor-1alpha (HIF-1alpha); pVHL30 is associated with microtubules. VHL exert its oncogenetic action both directly and through HIF-1alpha activation. TNM classification is unable to define a correct prognostic evaluation of intracapsular cc-RCC. The nucleo-cytoplasmic trafficking in VHL/HIF-1alpha pathway could be relevant in understanding the molecular pathogenesis of renal carcinogenesis. This study analyzes VHL/HIF-1alpha proteins in a large series of intracapsular cc-RCCs, correlating their expression and cellular localization with prognosis. MATERIALS AND METHODS: Two anti-pVHL (clones Ig32 and Ig33) and 1 anti-HIF-1alpha were used on tissue microarrays from 136 intracapsular cc-RCCs (mean follow-up: 74 mo). Clone 32 recognizes both pVHLs, whereas clone 33 only pVHL30. Results were matched with clinicopathologic variables and tumor-specific survival (TSS). RESULTS: A strong cytoplasmic positivity was found for all antibodies in the largest part of cases, associated to a strong nuclear localization in the case of HIF-1alpha. All pVHL-negative cases were associated with high HIF-1alpha expression. pVHL negativity and HIF-1alpha nuclear positivity significantly correlated with shorter TSS. In multivariate analysis both pVHL negativity and HIF-1alpha nuclear expression were independent predictors of TSS. CONCLUSIONS: The localization of the proteins well matches with their role and with the supposed tumor molecular pathways. The correlation with prognosis of VHL/HIF-1alpha alterations confirms the relevance of their molecular pathway and of the cellular trafficking of their products in the pathogenesis of renal cancer.     

Genetic evaluation of von Hippel–Lindau disease for early diagnosis and improved prognosis

von Hippel-Lindau disease (VHL) is a rare autosomal-dominant disorder in which affected individuals develop tumors in a number of locations. It occurs at a frequency of one per 36,000 population. Metastatic renal cell carcinoma (RCC) remains the leading cause of mortality in patients with clear cell RCC arising from mutations in the VHL tumor suppressor. RCC is the presenting feature in only 10% of VHL patients. VHL patients can present with a number of other renal lesions, such as hemangiomas and benign adenomas, in addition to simple cysts and RCC. We have investigated VHL gene mutations in familial RCC. The study cohort consisted of four patients with synchronous VHL and RCC and 31 kindreds. Analysis of the chromosomes was performed by the Moorehead method. Although none of the kindreds investigated had clinical evidence of VHL disease, 22 were found to have a VHL gene mutation consisting of deletions on the short arm of chromosomes 3, 17, and 19. Detailed clinical examination of the 22 kindreds with a VHL mutation revealed cerebellar hemangioblastoma (three kindreds), meningioma (two) and renal cell carcinoma (five). No VHL gene mutation was detected in nine kindreds. The prevalence of VHL gene mutations was 70.9% in the familial RCC kindreds. As a result of this study, the kindreds of patients with synchronous VHL and RCC have undergone molecular genetic testing and should be investigated for associated disorders.     

Somatic mutations of the von Hippel-Lindau disease gene in renal carcinomas occurring in patients with long-term dialysis.

BACKGROUND: Renal cell carcinoma (RCC) frequently occurs in patients with long-term dialysis. Long-term dialysis causes distinctive pathological changes in the kidney, which is known as acquired cystic disease of the kidney (ACDK). It is of great interest to know whether RCCs occurring in the dialytic kidneys harbour the same or similar mutations of the von Hippel-Lindau (VHL) gene as conventional dialysis-unrelated clear cell RCCs so often do. METHODS: Renal cancer tissues (eight clear cell, two papillary, one Bellini duct and three of the so-called dialysis-specific renal carcinomas) from 13 patients undergoing long-term dialysis were examined for somatic mutations of the VHL disease gene. By means of laser capture microdissection, cancerous and surrounding non-cancerous renal tissues from dialytic patients were subjected to PCR-based direct sequencing of the VHL gene. RESULTS: Direct forward and reverse sequencing showed that three tumours possessed VHL gene mutations (713delG, 500-504del5-bp and 709A>G). These three mutations were identified in clear cell carcinomas occurring in association with end-stage renal disease undergoing dialysis for 194, 147 and 125 months. None of the non-tumour tissues or other carcinoma tissues analysed, including dialysis-specific carcinoma, possessed VHL gene mutations. CONCLUSION: These results indicate that VHL tumour-suppressor gene mutation is involved in clear cell carcinoma in association with long-term dialysis. Mutation of the VHL gene was not found in any of the dialysis-specific RCCs studied herein.     

肾细胞癌VHL基因改变与VEGF表达的关系及意义

目的：探讨肾细胞癌VHL基因异常与血管内皮生长因子（VEGF）表达的关系及意义。方法：应用聚合酶链反应（PCR）加单链构象多态性分析（SSCP）、多聚合酶链反应（Multiplex-PCR)及免疫组织化学方法检测42例肾细胞癌、18例远离肿瘤的正常肾脏及10例正常肾脏组织中VHL基因突变、异常甲基化及VEGF的表达。结果：肾癌组织中VHL基因改变（61．9％）与正常肾脏组织（3．6％）比较差别有显著性意义（P＜0．005）,VHL基因失活与肾癌组织类型及临床分期相关,与病理分级无关。VEGF在肾癌组织（64．3％）与正常肾组织（21．4％）中的表达差别有显著性意义（P＜0．005）,肾癌组织中VEGF表达与组织类型无关,与病理分级及临床分期相关,随病理分级及临床分期的升高而增高。VHL基因改变与VEGF表达间存在显著的相关性（P＜0．05）。结论：VHL基因在肾细胞癌中具有高频突变率,可负向调节VEGF的表达。     

上海仁济医院肾癌数据库资料分析

目的 探讨肾癌临床、病理、分期、分级与预后特征. 方法 分析2003年至2005年上海仁济医院泌尿科肾癌数据库435例患者临床和病理资料.采用WHO 1997年肾实质上皮性肿瘤组织学分类标准、2002年ATCC的TNM分期和临床分期、1982年Fuhrman病理分级.采用Kaplan-Meier法和Logrank检验对57例获随访的晚期患者行生存分析和预后因素判断. 结果 435例患者中,遗传性VHL病肾癌10例(2.4%)、散发性肾透明细胞癌372例(85.5%)、乳头状癌13例(3.0%)、嫌色细胞癌18例(4.1%)、集合管癌4例(0.9%)、嗜酸性细胞腺瘤4例(0.9 %)、未分类肾癌.14例(3.2%).行根治性肾切除术335例(77.0%),保留肾单位手术74例(17.0%),姑息性肾切除等手术26例(6.0%).遗传性VHL病肾癌均为双肾癌伴多发囊肿,临床分期Ⅰ期7例、Ⅱ期3例,病理分级Ⅰ级6例、Ⅱ级4例,基因测序均存在VHL基因突变,平均随访28.6个月,患者无肿瘤局部进展或转移,但4例患者出现同侧或双侧肿瘤再发.嫌色细胞癌临床分期均为Ⅰ期,病理分级Ⅰ级5例,Ⅱ级13例,平均随访19.8个月均存活,无肿瘤转移或复发.集合管癌临床分期均为Ⅰ期,病理分级均为Ⅲ级,平均生存时间11.3个月.肾透明细胞癌和乳头状癌临床分期Ⅰ期260例(67.6%)、Ⅱ期64例(16.6%)、Ⅲ期32例(8.3%)、Ⅳ期29例(7.5%),其中T1a 147例(38.2%)、T1b 113例(29.4 %);病理分级Ⅰ级124例(32.2%)、Ⅱ级219例(56.9%)、Ⅲ级40例(10.4%)、Ⅳ级2例(0.5%).57例晚期肾癌患者中位生存时间(16.0±1.3)个月,1年生存率55.0%,2年生存率31.0%.预后因素分析显示,临床分期、肿瘤大小、淋巴结转移、远处转移和病理分级是晚期肾癌解剖水平和组织学水平的预后影响因素. 结论 不同组织学亚型的肾癌生物学特征存在较大差异,遗传性VHL病肾癌存在基因突变,常为双侧、多中心、低Fuhrman分级透明细胞癌,易再发不易转移.肾嫌色细胞癌预后较好,而集合管癌预后差.在解剖水平和组织学水平,TNM分期、肿瘤大小、淋巴结转移、远处转移和肾癌病理分级是晚期肾癌的预后影响因素.